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Enzyme Kinetics Lab Packet (1).docx - Brigham Young...

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Brigham Young University – Idaho Bio 180 Lab Lab 5: Enzyme Kinetics Objectives 1. To examine the influence of temperature on the activity of the enzyme peroxidase Introduction Chemical reactions occurring in living organisms are carried out by biological catalysts called enzymes. Enzymes are proteins made up of amino acids held together by strong covalent bonds called peptide bonds. The sequence of amino acids (determined by the DNA sequence) ultimately determines the structure and function of the enzyme. Polypeptides fold into 3-dimensional structures in order to become functional and carry out their catalytic reactions. This 3-dimensional folding occurs when weak bonds form between amino acids. These weak interactions can be affected by the environment in which enzymes reside, and any change in the 3-dimensional shape of the enzyme may alter its ability to function normally. Enzymes contain active sites, the region where substrates (reactants) attach and where the chemical reaction takes place. Active sites are highly specific and will bind to some substrates but not to others. The reaction is complete when the end products are formed and released from the active site. The enzyme can now undergo another catalytic reaction with a new substrate. In summary, environmental conditions affect chemical reaction rates by changing the shape of the enzyme and its active site. Kinetics is the study of the rates of chemical reactions. The range of pH, temperature, salt concentration, and substrate concentration in a cell all contribute to the activity of an enzyme, such that the conditions under which the enzyme functions at its maximal capacity are called the optimal conditions. Optimal conditions will be different for every enzyme in one organism, as well as for the same enzyme in organisms of different species. The enzyme that will be studied today in lab is peroxidase. Peroxidase is found in peroxisomes, where it converts toxic hydrogen peroxide into harmless water and molecular oxygen: 2 H 2 O 2 2 H 2 O + O 2 The activity of this enzyme can be followed by measuring the amount of oxygen liberated in the reaction. Some dyes will react with oxygen and change from a colorless to a colored state. One of these dyes is Guaiacol.
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